Device and method for the controlled heating in micro channel systems

ABSTRACT

A method of controlled heating of a micro channel reactor structure ( 46, 48, 50 ) comprises providing a structure (b 1 , b 2 , B 1 , B 2 ) defining a desired temperature profile. A preferred embodiment of a heating element structure comprises a pattern of areas of a material capable of providing heat when energized, disposed over said micro channel reactor structure.

The present invention relates to methods and devices for the controlledheating, in particular of liquid samples in small channels that arepresent within a substrate.

BACKGROUND OF THE INVENTION

There is a trend in the chemical and biochemical sciences towardsminiaturization of systems for performing analytical tests and forcarrying out synthetic reactions, where large numbers of reactions mustbe performed. For example in screening for new drugs as many as 100000different compounds need to be tested for specificity by reacting withsuitable reagents.

Another field is polynucleotide amplification, which has become apowerful tool in biochemical research and analysis, and the techniquestherefor have been developed for numerous applications. One importantdevelopment is the miniaturization of devices for this purpose, in orderto be able to handle extremely small quantities of samples, and also inorder to be able to carry out a large number of reactions simultaneouslyin a compact apparatus.

In most systems for the purposes indicated above (and others notmentioned) there would commonly be a need for heating the reagents insome stage of the procedure for carrying out the necessary reactions.Even more importantly there is also a need for maintaining the reactiontemperature at a constant level during a desired period of time, i.e. toavoid variations in temperature across the channel part containing thereagents that have been heated (reactor volume).

Furthermore, in these miniaturized systems the temperature of the samplewill essentially be determined by the temperature of the wall confiningthe sample. Thus, if the material constituting the wall leads away heat,there will be a temperature drop close to the wall, and a variationthroughout the sample occurs.

There is also a problem with evaporation when heating small aliquots ofliquids within micro channel structures. This problem can be solved byproviding heating means in the form of a surface layer that is capableof absorbing light energy for transport into a selected area See WO0146465 (FIG. 7 and related disclosure). Conveniently white light isused, but for special purposes, monochromatic light (e.g. laser) canalso be used. The layer can be a coating of a light-absorbing layer, e.ga. black paint, which converts the influx of light to heat.

An alternative solution to the evaporation problem has been to carry outthe steps involving elevated temperature (heating steps) within closedreaction volumes. This has required solving problems related the largepressure increase that typically is at hand when heating liquid aliquotswithout venting. If the process concerned is integrated into a sequenceof reactions there is a demand for smart valving solutions.

In many of the prior art devices the substrate material has had a fairlyhigh thermal conductivity which has permitted heating by ambient air orby separate heating elements in close association with the inner wall ofthe channel containing a liquid to be heated. Cooling has typicallyutilized ambient air. Recently it has become popular to manufacturemicro channel structures in plastic material that typically has a lowthermal conductivity. Due to the poor thermal conductivity, unfavorabletemperature gradients may easily be formed within the selected area whenthis latter type of materials is used. These gradients may occur acrossthe surface and downwards into the substrate material. The variation intemperature may be as high as 10° C. or more between the center of thearea or region and its peripheral portions. If the light absorbing areais too small this variation will be reflected in the temperature profilewithin a selected area and also within the heated liquid aliquot. Formany chemical and biochemical reactions such lack of uniformity can bedetrimental to the result, and indeed render the reaction difficult tocarry out with an accurate result.

Although the heating means according to WO 0146465 eliminates theevaporation and the pressure problem, it still suffers from theabove-mentioned temperature variation across the sample. Suchtemperature variations are often detrimental to the outcome of areaction and must be avoided.

Microfluidic platforms that can be rotated comprising heating elementshave been described in WO 0078455 and WO 9853311. These platforms areintended for carrying out reactions at elevated temperature, forinstance thermal cycling.

SUMMARY OF THE INVENTION

In view of the shortcomings of prior art systems, it would be desirableto have access to a device for performing chemical/biochemicalreactions/analyses, such as but not limited to, polynucleotideamplification reactions, in which controlled heating of the reactants ina small reaction volume, e.g. a capillary, can be performed withoutcausing the uncontrolled evaporation discussed above, and where thetemperature can be maintained at a constant level throughout thereaction volume. The object of the invention is thus to accomplish aproper balance between influx of heat and cooling so that a liquidaliquot in a micro channel can be quickly heated and maintained at auniform temperature for well defined time intervals.

The above indicated object can be achieved in accordance with thepresent invention by a method of controlled heating as claimed in claims1–10, and a micro channel reactor system as claimed in claims 11–20. Infurther aspects the invention provides a heating structure, as claimedin claim 21–26, a rotatable disc as claimed in claims 27–29. In apreferred embodiment the system is implemented by employing a rotatingmicrofluidic disc. Such devices employ centrifugal force to drive sampleand reagent through the system of channels and reaction chambers.Spinning assists in establishing the proper heat balance to maintain auniform temperature within the reactor.

In the context of the invention the term “selected area” means theselected surface area to be heated plus the underlying part of thesubstrate containing the reactor volume of one or more micro channels ifnot otherwise being clear from the particular context. The selected areacontains substantially no other essential parts of the micro channels.The term “surface” will refer to the surface to be heated, e.g. thesurface collecting the heating irradiation, if not otherwise indicated.

By the terms “heating structure”, “heating element structure” and“heating element” are meant a structure which is present in or on aselected area, or between the substrate and a radiation source, andwhich defines a pattern which (a) covers a selected area and (b) can beselectively heated by electromagnetic radiation or electricity, such aswhite or visible light or only IR, or by direct heating such aselectricity. In this context the term “pattern” means (1) a continuouslayer, or (2) a patterned layer comprising one or more distinct partsthat are heated and one or more distinct parts that are not heated. (b)excludes that the pattern consists of only the part that is heated.

A preferred variant of a heating structure is given in claims 21–26.

The invention will now be described in detail with reference to theattached drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1 a–d illustrates a prior art microfluidic disc;

FIGS. 2 a–b illustrates a prior art device with (a) a heating structureand (b) a temperature profile across the selected area during heating;

FIGS. 3 a–c illustrates the difference between (a) a prior art surfacetemperature profile and (b) a desired surface temperature profileaccording to the invention, and a typical temperature profile betweenthe opposing surfaces of a selected area made of plastic material;

FIGS. 4 a–e exemplifies various micro channel structures to which theinvention is applicable;

FIGS. 5 a–b illustrates a microfluidic disc and an embodiment of aheating element structure according to the invention;

FIGS. 6 a–b illustrates a further embodiment of a heating elementstructure and the obtainable temperature profile;

FIGS. 7 a–c illustrates still another embodiment of a reactor system andan inventive heating element structure and the obtainable temperatureprofile;

FIGS. 8 a–c is a further embodiment implemented for another geometry;

FIGS. 9 a–b are embodiments of a resistive heating element structureaccording to the invention; and

FIGS. 10 a–b illustrates means for controlling the flanks of thetemperature profile.

DETAILED DESCRIPTION OF THE INVENTION

For the purpose of this application the term “micro channel structure”as used herein shall be taken to mean one or more channels, optionallyconnecting to one or more enlarged portions forming chambers having alarger width than the channels themselves. The micro channel structureis provided beneath the surface of a flat substrate, e.g. a disc member.

The terms “micro format”, “micro channel” etc contemplate that the microchannel structure comprises one or more chambers/cavities and/orchannels that have a depth and/or a width that is ≦10³ μm preferably≦10² μm. The volumes of micro cavities/micro chambers are typically≦1000 nl, such as ≦500 nl or ≦100 nl or ≦50 nl. Chambers/cavitiesdirectly connected to inlet ports may be considerably larger, e.g. whenthey are intended for application of sample and/or washing liquids.

In the preferred variants volumes of the liquid aliquots used are verysmall, e.g. in the nanoliter range or smaller (≦1000 nl). This meansthat the spaces in which reactions, detections etc are going to takeplace often becomes more or less geometrically indistinguishable fromthe surrounding parts of a micro channel.

A reactor volume is the part of a micro channel in which the liquidaliquot to be heated is retained during a reaction at an elevatedtemperature. Typically reaction sequences requiring thermal cycling orotherwise elevated temperature take place in the reaction volume. Thedisc preferably is rotatable by which is meant that it has an axis ofsymmetry (C_(n)) perpendicular to the disc surface. n is an integer 3,4, 5, 6 or larger . The preferred discs are circular, i.e. n=∞. A discmay comprise ≧10 such as ≧50 or ≧100 or ≧200 micro channels, each ofwhich comprising a cavity for thermo cycling. In case of discs that canrotate, the micro channels may be arranged in one or more annular zonessuch that in each zone the cavities for thermo cycling are at the sameradial distance. By the expressions “essentially uniform temperatureprofile” and “constant temperature” are meant that temperaturevariations within a selected area of the substrate are within suchlimits that a desired temperature sensitive reaction can be carried outwithout undue disturbances, and that a reproducible result isachievable. This typically means that within the reaction volume, thetemperature varies at most 50%, such as at most 25% or at most 10% or 5%of the maximum temperature difference between the opposing surfaces ofthe selected area comprising the heated liquid aliquot. These permittedvariations apply across a plane that is parallel to the surface and/oralong the depth of the micro channel. The acceptable temperaturevariation may vary from one kind of reaction to another, although it isbelieved that the acceptable variation normally is within 10° C., suchas within 5° C. or within 1° C.

The present invention suitably is implemented with micro channelstructures for a rotating microfluidic disc of the kind, but not limitedthereto, disclosed in WO 0146465, and in FIG. 1 in the presentapplication, there is shown a device according to said application.However, it is to be noted that this is only an example and that thepresent invention is not limited to use of such micro channelstructures.

The micro channel structures K7–K 12 according to this known device,shown in FIGS. 1 a–d, are arranged radially on a microfluidic disc D.Suitably the microfluidic disc is of a one- or two-piece mouldedconstruction and is formed of an optionally transparent plastic orpolymeric material by means of separate mouldings which are assembledtogether (e.g. by heating) to provide a closed unit with openings atdefined positions to allow loading of the device with liquids andremoval of liquid samples. See for instance WO 0154810 (Gyros AB).Suitable plastic of polymeric materials may be selected to havehydrophobic properties. In the alternative, the surface of the microchannels may be additionally selectively modified by chemical orphysical means to alter the surface properties so as to producelocalised regions of hydrophobicity or hydrophilicity within the microchannels to confer a desired property. Preferred plastics are selectedfrom polymers with a charged surface, suitably chemically or ion-plasmatreated polystyrene, polycarbonate or other rigid transparent andnon-transparent polymers (plastic materials). The term “rigid” in thiscontext includes that discs produced from the polymers are flexible inthe sense that they can be bent to a certain extent. Preferred plasticmaterials are selected from polystyrenes and polycarbonates. In case theprocess taking place within the micro channel structure requires opticalmeasurement, for instance of fluorescence, the preferred plasticmaterials are based on monomers only containing saturated hydrocarbongroups and polymerisable unsaturated hydrocarbon groups, for instanceZeonex® and Zeonor®. Preferred ways of modifying the plastics by plasmaand by. hydrophilization are given in WO 0147637 (Gyros AB) and WO0056808 (Gyros AB).

The micro channels may be formed by micro-machining methods in which themicro-channels are micro-machined into the surface of the disc, and acover plate, for example, a plastic film is adhered to the surface so asto close the channels. Another method that is possible is injectionmolding. The typical microfluidic disc D has a thicknes, which is muchless than its diameter and is intended to be rotated around a centralhole so that centrifugal force causes fluid arranged in the microchannels in the disc to flow towards the outer periphery of the disc. Inthe embodiment of the present invention shown in FIGS. 1 a–1 d, themicro channels start from a common, annular inner application channel 1and end in common, annular outer waste channel 2, substantiallyconcentric with channel 1. It is also possible to have individualapplication channels (waste channels for each micro channel or a groupof micro channels). Each inlet opening 3 of the micro channel structuresK7–K12 may be used as an application area for reagents and samples. Eachmicro channel structure K7–K12 is provided with a waste chamber 4 thatopens into the outer waste channel 2. Each micro channel K7–K12 forms aU-shaped volume defining configuration 7 and a U-shaped chamber 10between its inlet opening 3 and the waste chamber 4. The normal desiredflow direction is from the inlet opening 33 to the waste chamber 4 viathe U-shaped volume-defining configuration 7 and the U-shaped chamber10. Flow can be driven by capillary action, pressure, vacuum andcentrifugal force, i.e. by spinning the disc. As explained later,hydrophobic breaks can also be used to control the flow. Radiallyextending waste channels 5, which directly connect the annular innerchannel 1 with the annular outer waste channel 2, in order to remove anexcess fluid added to the inner channel 1, are also shown.

Thus, liquid can flow from the inlet opening 3 via an entrance port 6into a volume defining configuration 7 and from there into a first armof a U-shaped chamber 10. The volume-defining configuration 7 isconnected to a waste outlet for removing excess liquid, for example,radially extending waste channel 8 which waste channel 8 is preferablyconnected to the annular outer waste channel 2. The waste channel 8preferably has a vent 9 that opens into open air via the top surface ofthe disk. Vent 9 is situated at the part of the waste channel 8 that isclosest to the centre of the disc and prevents fluid in the wastechannel 8 from being sucked back into the volume-defining configuration7.

The chamber 10 has a first, inlet arm 10 a connected at its lower end toa base 10 c, which is also connected to the lower end of a second,outlet arm 10 b. The chamber 10 may have sections I, II, III, IV whichhave different depths, for example each section could be shallower thanthe preceding section in the direction towards the outlet end, oralternatively sections I and III could be shallower than sections II andIV, or vice versa A restricted waste outlet 11, i.e. a narrow wastechannel is provided between the chamber 10 and the waste chamber 4. Thismakes the resistance to liquid flow through the chamber 10 greater thanthe resistance to liquid flow through the path that goes throughvolume-defining configuration 7 and waste channel 8.

By introducing a well defined volume of sample that will just about fillone U shaped volume of this configuration, it will be possible toconfine this sample within the portion of the micro channel structurethat is defined by said U, by spinning the disc, and thus impose asimulated gravity. If the spinning speed is sufficient, the forceimposed will force the condensed droplets back into the reaction volume.If heating is applied locally and the material of the disc has a lowthermal conductivity, for instance plastics, a steep decreasingtemperature gradient will form between the heated and non-heated area.The upper part of the arms will act as a cooler and assist incounteracting evaporation. The need for securing evaporation losses byclosing the system can be avoided. Thus, in fact the U shaped volumewill be an effective reaction chamber for the purpose of thermalcycling, e.g. for performing polynucleotide amplification by thermalcycling.

The term “U-shaped” includes also other shapes in which the channelstructure comprises a bent towards the periphery of the disc and twoinwardly directed arms, for instance Y-shaped structures where thedownward part is pointing towards the periphery of the disc andcomprises a valve function that is closed while heating at least thelower part of the upwardly directed arms.

However, it is equally possible to use a micro channel structure withoutthe above discussed U-configuration, namely by employing a straight,radially extending channel, but provided with a stop valve at the endclosest to the disc circumference. A valve suitable for this purpose isdisclosed in SE-9902474-7, the disclosure of which is incorporatedherein in its entirety.

Such a valve operates by using a plug of a material that is capable ofchanging its volume in response to some external stimulus, such aslight, heat, radiation, magnetism etc. Thus, by introducing a sample ina capillary at a desired location, sealing the capillary at theoutermost end position of the sample, and spinning the disc, the samplewill be held in place, and uncontrolled evaporation during heating canbe controlled in the same way as in the embodiment employing aU-configuration.

Also mechanical valves can be used in the variants mentioned above.

However, as indicated above, it is essential that a uniform temperaturelevel can be maintained locally in the entire reaction volume preferablywith a steep temperature gradient to the non-heated parts of themicrofluidic substrate. Such controlled heating is convenientlyperformed by a heating system and method according to the presentinvention, embodiments of which will now be described in detail below.The heating system referred to in this paragraph may be based on contactheating or non-contact heating.

FIG. 2 a shows a micro channel structure having a U configuration 20provided on a microfluidic disc of the type discussed previously, whichis covered by a light absorbing area 22 for the purpose of heating. FIG.2 b shows a temperature profile across said light absorbing area alongthe indicated centerline b—b, when it is illuminated with white lightlight. As can be clearly seen, the temperature profile is bell shaped,which unavoidably will cause uneven heating within the region where thechannel structure is provided, thus causing the chemical reactions torun differently in said channel structure at different points.

It would be possible to enlarge the area such that its periphery islocated sufficiently remote from the channel structure that the bellshaped temperature profile is “flattened” out to an extent that therewill be a more uniform temperature across the reactor volume. However,in the first place this would require too much surface around thechannel structure to be covered by the light-absorbing layer, and sincethere is a desire to provide a very large number of channel structuresclose to each other, an enlarged area would occupy too much surface.Secondly, even if a very large area is provided the temperature profilewould still exhibit a more or less clear bell shape, indicatingnon-uniform temperature over the channel structure defining the reactionvolume.

In essence, it all comes down to enabling heating of a local area of asubstrate containing a micro channel/chamber structure, in a controlledway, so as to achieve a uniform heating across the volume containing theliquid aliquot to be heated. This should be achieved at the same time assurrounding elements and materials should be as little affected aspossible by the heating, i.e. preferably, areas immediately adjacent theheated region, e.g. comprising another part of the micro channelstructure, should not be heated at all, in the ideal situation. It is ofcourse desirable that the temperature is equal throughout the entirevolume. In the case of the present invention implemented in small microchannels and heating at the surface closest to the micro channel, theinventive heating method and heating element structure, primarilyensures a uniform temperature level in the sense as defined above to beachieved across the surface of a selected area where the microchannel(s) is (are) located. The factual variations that may be at handin the surface becomes smaller in any plane inside the selected area.The plane referred to is parallel with the surface. However, there willbe a relatively large temperature drop through the thickness of thedisc. This drop is typically of the order of 10° C. In spite of this,because the channel dimensions are so small, only about 1/10 of thethickness of the substrate, the temperature drop over the channel inthis direction will be only about 1° C., which is acceptable for allpractical purposes. This is illustrated in FIG. 3 c. This relativelylarge temperature drop along the thickness of the substrate will assistin an efficient and rapid cooling of the heated liquid aliquot after aheating step. This becomes particularly important if the processperformed comprises repetitive heating and cooling (thermal cycling) ofthe liquid aliquot. Cooling will be assisted by spinning the disc.

When a disc is rotated, the frictional forces will drag air at thesurface of the disc. Thus, the air near the disc will rotate in the samedirection as the disc. The rotation of the air will result incentrifugal forces that will cause the air to flow radially.

The flowing air will have a cooling effect on the surface of the disc,and in fact it is possible to control the rate of cooling veryaccurately by controlling the speed of rotation, given that the airtemperature is known. This effect is utilized in the present invention,and is a key factor for the success of the heating method and systemaccording to the invention.

It would be possible to obtain the same effect if one uses controlledair flow from a fan or the like, where the cooling effect can be variedby varying the speed of the fan. This method could be used forstationary systems where the regions, e.g. comprising micro channelstructures, to be cooled are made in e.g. a flat substrate, which isnon-rotary.

Most plastic materials, in particular transparent plastic materials arenon-absorbing with respect to visible light but not to infrared. Formicrofluidic discs made of transparent polymeric materials, illuminationwith visible light will cause only moderate heating (if any at all),since most of the energy is not absorbed. One possibility to convertvisible light to heat in a defined area or volume (selected area) is toapply a light absorbing material at the location where heating isdesired.

Thus, in order to transform light to heat, such light absorbing materialmust be provided at the position where heating is desired. This canconveniently be achieved by covering the position or region with e.g.black color by printing or painting. When illuminated, the lightabsorbing material will become warm, and heat is transferred to thesubstrate on which it is deposited. Between the various spots of lightabsorbing material there may be a material reflecting the irradiationused. An alternative for the same kind of substrates is to cover one ofthe substrate surfaces with a light absorbing material and illuminatingthis surface through a mask only permitting light to pass through holesin the mask that are aligned with the selected areas.

For substrates made in plastic material that absorbs the radiation used,the surface may be coated with a mask that reflects the radiationeverywhere except for the selected areas. Alternative the mask may bephysically separated from the substrate but still positioned between thesurface of the substrate and the irradiation source.

In accordance with the present invention, the area is given a specificlay-out that changes the temperature profile, from a bell shape to(ideally) an approximate “rectangular” shape, i.e. making thetemperature variation uniform across the surface of the selected area oracross a plane parallel thereto. One method is by a simple trial enderror approach. For non-absorbing materials, a pattern of materialabsorbing the radiation is placed between the surface of the substrateand the source of radiation. Typically the material is deposited on thesubstrate. By using an IR video camera the temperature at the surfacecan be monitored. Another method for arriving at said layout is byemploying FEM calculations (Finite Element Method). FIG. 3 illustratesschematically the change in profile principally achievable by employingthe inventive idea The bell shaped profile A results with a lightabsorbing area A having the general extension as shown FIG. 3 a, (theprofile taken in the cross section indicated by the arrow a), and the“rectangular” profile results when employing a light absorbing region asshown by curve B in FIG. 3 (the profile taken in the cross sectionindicated by the arrow).

The most important feature of the temperature profile is that its upper(top) portion is flattened (uniform), thus implying a low variation intemperature across the corresponding part of the selected area. The“flanks”, i.e. the side portions of the profile will always exhibit aslope, but by suitable measures this slope can be controlled to theextent that the profile better will approximate an ideal rectangularshape.

Now various embodiments of the present invention and different aspectsthereof will be described with reference to the drawings.

In a first embodiment of the invention, electromagnetic radiation, forinstance light, is used for heating a liquid present in a selected areaof a substrate made of a plastic material not absorbing the radiationused for heating. In this case a surface of the selected area iscovered/coated with a layer absorbing the radiation energy, e.g. light.As outlined in this specification the kind of radiation, plasticmaterial and absorbing layer must match each other. The layer may be ablack paint. The paint is laid out in a pattern of absorbing andnon-absorbing (coated and non-coated) parts (subareas) on the surface ofthe selected areas. The term “non-absorbing part” includes covering witha material reflecting the radiation. In other variants of thisembodiment, the layer absorbing the irradiation is typically within thesubstrate containing the micro channel. In the case quick and/orrelatively high increase in temperature is needed, the distance betweenthe layer absorbing the irradiation used and reactor volume at most thesame as the shortest distance between the reactor volume and the surfaceof the substrate. A relatively high increase in temperature means up tobelow the boiling point of water, for instance in the interval 90–97° C.and/or an increase of 40–50° C. The absorbing layer may also located tothe inner wall of the reactor volume.

The first embodiment also includes a variant in which the substrate ismade of plastic material that can absorb the electromagnetic radiationused. In this case a reflective material containing patterns ofnon-absorbing material including perforations is placed between thesurface of the selected areas and the source of radiation. This includesthat the reflective material for instance is coated or imprinted on thesurface of the substrate. Non-adsorbing patterns, for instance patternsof perforation, are selectively aligned with the surfaces of theselected areas. This variant may be less preferred because absorption ofirradiation energy will be essentially equal throughout the selectedarea that may counteract quick cooling.

By the term “absorbing plastic material” is meant a plastic materialthat can be significantly and quickly heated by the electromagneticradiation used. The term “non-adsorbing plastic material” means plasticmaterial that is not significantly heated by the electromagneticradiation used for heating.

The term “pattern” above means the distribution of both absorbing andnon-absorbing parts (subareas) across a layer of the selected area, forinstance a surface layer. The term excludes variants where the patternonly comprises one absorbing part covering completely the surface of theselected area.

The invention will now be illustrated by different patterns of absorbingmaterials coated on substrates made of non-absorbing plastic material.For substrates made of absorbing plastic material, similar patternsapply but the non-absorbing parts are replaced with a reflectivematerial and the absorbing parts are typically uncovered.

As a first example let us consider a micro channel/chamber structure, afew examples of which are indicated in FIGS. 4 a–e. This kind ofchannel/chamber structures can be provided in a large number, e.g. 400,on a microfluidic disc 40 (schematically shown in FIG. 5 a). Allchannel/chamber structures need not be identical, but in most cases theywill be, for the purpose of carrying out a large number of similarreactions at the same time. If we assume that all channel/chamberstructures are identical, and that only one portion (e.g. a reactionchamber or a segment of a channel) of the channel/chamber structureneeds to be heated during the operation, it will be convenient toprovide the inventive heating element structure, e.g. as in FIG. 3 b, asconcentric bands of paint 42, 44, as shown in FIG. 5 b, or some otherkind of absorbing material.

The provision of this basic band configuration is not an optimalsolution, however, since the temperature profile still exhibits a slightfluctuation over the area to be heated. In a preferred embodimenttherefore, there is provided several narrow bands b1, b2 of lightabsorbing material (paint) between the larger bands B1, B2, asschematically shown in FIG. 6 a, which shows a broken away view of adisc 40 having a plurality of channel structures 46, 48, 50. In FIG. 6 bthe corresponding temperature profile achievable with this bandconfiguration is shown. In this example it is the part of the microchannel structure delimited by the square A (FIG. 6 a) that it isdesired to heat in a controlled manner.

The heating element structure described above is applicable to allchannel/chamber structures shown in FIG. 4.

However, for certain applications it can be desirable to provide evenmore localized heating, e.g of a circular or rectangular/square area.This would especially be required if adjacent or surrounding areas mustnot be heated at all. The embodiment with concentric bands of paint willresult in heating also of the areas between the radially extending microchannel/chamber structures.

In FIG. 7 a there is shown a micro channel/chamber structure 70 with acircular chamber with an inlet 71 and an outlet 72 channel. If it isimportant to avoid heating of the disc area surrounding the chamber, aheating element structure as shown in FIG. 7 b can be employed,comprising concentric bands B1, b2 and a center spot c1. In this casethe temperature profile will be the same in all cross sections throughthe center of the micro channel/chamber structure, and look somethinglike the profile of FIG. 7 c.

In FIGS. 8 a–c a similar channel structure, but applied to a rectangularchamber is shown. FIG. 8 c shows the temperature profiles C1, C2 indirections c1 and c2 of FIG. 8 b, respectively.

For the illumination, lamps of relatively high power is used, suitablye.g. 150 W. Suitable lamps are of the type used in slide projectors,since they are small and are provided with a reflector that focuses theradiation used. The irradiation can be selected among UV, IR, visiblelight and other forms of light as long as one accounts for matching thesubstrate material and the absorbing layer properly. In case the lampgives a desired wave-length band but in addition also wavelengths thatcause heat production within the substrate it may be necessary toinclude the appropriate filter. In order to achieve the best results thelight should be focussed onto the substrate corresponding to a limitedregion on the substrate, e.g. about 2 cm in diameter, although of coursethe size may be varied in relation to the power of the lamp etc. One ormore lamps could be used in order to enable illumination of one or moreregions, e.g. in the event it is desirable to carry out differentreactions at different locations on a substrate On a rotating disc itmight be desirable to perform heating at different radial locations.Illumination of the substrate can be from both sides. If the lightabsorbing material is deposited on the bottom side, nevertheless theillumination can be on the topside, in which case light is transmittedthrough the substrate before reaching the light absorbing material.Illumination of the backside with material deposited on the topside isalso possible.

In view of the spinning speed of a rotating microfluidic disc being ashigh as of the order of 1000 rpm, the pulsing effect obtained in thisway will not be noticeable and the heating can for all practicalpurposes be considered as continuous.

The above described embodiments have employed light absorbing materialto provide the heating elements, but it is within the scope of theinvention to employ any heating element structure in a suitable patternthat is capable of generating heat. Thus, it is also contemplated toprovide areas of a resistive material 91, 92 in the same general layoutsas shown in FIGS. 7–8. Examples thereof applied to the same channelstructures as those in FIGS. 7–8 are shown in FIGS. 9 a–b.

The patterns are applied e.g. by printing of ink comprising conductiveparticles, e.g. carbon particles mixed with a suitable binding agent,using e.g. screen printing techniques. Patterns functioning in the sameway may also be created by the following steps

-   -   (a) covering the surface of a substrate made of non-absorbing        material with absorbing material and    -   (b) placing a reflective mask which contains patterns of holes        or of non-absorbing material between the surface of the        substrate and the source of the radiation with the individual        patterns being aligned with the surfaces of the selected areas.

Another aspect that should be considered for the performance is theeffect of cooling from the air flowing on the disc when it is rotated.Let us consider the configuration shown in FIG. 6 again. By the spinningaction air will be forced radially outwards over the surface of the discand will thereby cool the surface by absorbing some heat, such that theair is also heated. Thus, the air temperature will be higher towards theperiphery of the disc, and the non-coated (non-painted) area between thebands of light absorbing material nearest the periphery will thereforenot be as efficient in terms of decreasing the temperature as thenon-coated/non-absorbing area between the bands of light absorbingmaterial closer to the center.

In order to compensate for this phenomenon, the width of the non-coatedareas can be larger nearer the periphery than the width of those nearerthe center.

Normally the rotatable disc comprises a base portion having a top and abottom side, on the top side of which said micro channel structure isprovided, and on top of which a cover is provided so as to seal themicro channel structure. The heating elements (layer absorbing radiationenergy) are preferably provided on the top surface so as to cover theselected area to be heated. However, said light absorbing layer canalso, as an alternative, be provided on said bottom side.

In still another embodiment the heating element structures according tothe invention can be applied to stationary substrates, i.e. chip typedevices. In case of stationary substrates it will be necessary to useforced convection, e.g. by using fans or the like to supply thenecessary cooling. In all other respects the micro channel/chamberstructures and heating structures can be identical.

As mentioned above the flanks of the temperature profile exhibits acertain slope, which has as a consequence that an area surrounding thepart of the micro channel structure that is to be heated, will also beheated. This is because the substrate material adjacent the region whichis coated will dissipate heat from the area beneath the coating. One wayof reducing this heat dissipation is to reduce the cross section forheat conduction. This can be done by providing a recess 93 in thesubstrate 94 on the opposite side of the coating 95 along the peripheryof said coating as shown in FIG. 10 a. In this way the resistance toheat being conducted away from the coated region will be increased.Another way to obtain a similar result is to provide holes 96 instead ofsaid recess, but along the same line as said recess, as shown in FIG. 10b.

In the present invention it is used to an advantage that the heatconductivity of the substrate material, e.g. polymer, is poor. Thus,when the reaction volume is heated by using the inventive heatingstructure, the heat will not easily dissipate into the surroundingregions. Therefore, when the reaction inside the heated volume takesplace and if/when evaporation of liquid in the reaction volume occurs,any vapors formed, striving to move upstream in the micro channelstructure, will experience a cooler part of the channel, and willrapidly condense to liquid. In the case of a rotating disc system, theimposed gravity will then force the liquid droplets back into thereaction volume, and thereby reaction conditions will be controlled interms of keeping the sample volume variation within acceptable limits(i.e. negligible or no loss of sample due to evaporation), and also theconcentration of sample will be controlled to a reasonable extent(solvent will reflux into the reaction volume). If a stationary chiptype system is used, pressure can be applied to force the condensedvapors to flow back into the reaction volume.

One further aspect of the invention is an instrument comprising arotatable disc as defined in any of claims 27–29 and a spinner motorwith a holder for the disc, said motor enabling spinning speeds that arepossible to regulate. Typically the spinning of the motor can beregulated within an interval that typically can be found within 0–20 000rpm. The instrumentation may also comprise one or more detectors fordetecting the result of the process or to monitor part steps of theprocess, one or more dispensers for introducing samples, reagents,and/or washing liquids into the micro channel structure of the substratetogether with means for other operations that are going to be performedwithin the instrument.

One additional aspect of the invention is a method for performing areaction at elevated uniform temperature in one or more reactionmixtures (liquid aliquots). This aspect is characterized in comprisingthe steps of:

-   -   (i) providing a rotatable microfluidic disc as defined in any of        claims 27–29;    -   (ii) introducing said one or more reaction mixtires into        separate reaction volumes in the microfluidic disc;    -   (iii) supplying energy to the heating structure of the        microfluidic disc to increase the temperature in the reaction        mixtures to said elevated temperature and maintaining the        temperature at the elevated level for a sufficient time for the        intended reaction to take place;    -   (iv) possibly reducing the temperature,    -   (v) transfering each reaction mixture further downstream in the        micro channel linked to the reactor volume in which the mixture        has been processed,        with the provision that at least steps (iii) and (iv) are        carried out while spinning the disc with the spinning speed        being higher during step (iv) compared to during step (ii)        and/or the energy input to the heating structure being lower        during step (iv) than during step (iii).

Although the invention has been described with reference to thedrawings, but it should not be regarded as limited to the shownembodiments, the scope of the invention being defined by the appendedclaims. Thus, modifications and variations beyond the illustratedexamples are within the scope of the claims.

1. A method of providing a uniform temperature profile across a reactionvolume that is part of a microchannel structure that is contained in asubstrate, comprising the steps of (i) providing a heating structuredefining a) a selected area to be heated on said substrate which areaincludes said reaction volume, and b) the uniform temperature profileacross said reaction volume; and said heating structure comprising amaterial provided on said substrate, the material being capable oftransferring heat into said selected area when suitably energized and islaid out in a pattern that is capable of causing heating and cooling tobalance each other so as to create said uniform temperature profilewithin said reaction volume; and (ii) supplying energy to the substrate,whereby the presence of said heating structure causes essentially onlythe selected area to be heated creating said uniform temperature profilewithin said reaction volume.
 2. The method as claimed in claim 1,further comprising providing cooling by a flow of air over saidsubstrate.
 3. The method as claimed in claim 1, wherein the substrate isa rotatable disc, said heating structure being capable of absorbingelectromagnetic energy, and wherein heat energy is supplied byirradiation of the disc using a source of electromagnetic radiation,wherein the source is a light source.
 4. The method as claimed in claim1, wherein heat energy is supplied by irradiation of the disc using asource of electromagnetic radiation, wherein said source is a lightsource and wherein said heating structure is provided by 1) a separatemask element, inserted between the substrate and said light source, and2) by a material covering the substrate, and being capable of absorbingelectromagnetic energy.
 5. The method as claimed in claim 3, comprisingspinning the disc and illuminating the disc, the light being focusedonto the substrate corresponding to a limited region on the substrate.6. The method as claimed in claim 2, wherein the substrate is astationary substrate.
 7. The method as claimed in claim 6, wherein thecontrolled flow of air is provided with a fan.
 8. The method as claimedin claim 2 further comprising changing the temperature by changing thespeed of rotation of said disc and/or by reducing the energy of theelectromagnetic radiation.
 9. A micro-channel reactor system forcreating and maintaining a uniform temperature profile within a selectedreaction volume in the reactor system, comprising i) a substrate havingat least one micro-channel structure, the micro-channel structurecomprising one or more micro-channels which includes the reactionvolume; and ii) a heating structure defining a) a selected area to beheated on said substrate which includes the reaction volume of one ormore microchannels, and b) the uniform temperature profile across saidreaction volume; wherein said heating structure comprises a materialprovided on said substrate, the material being capable of transferringheat into said reaction volume when suitably energized, said materialbeing provided on at least one side of said substrate, and being laidout in a pattern, that is capable of causing heating and cooling tobalance each other so as to create said uniform temperature profile insaid reaction volume.
 10. The reactor system as claimed in claim 9,wherein said substrate is a rotatable disc.
 11. The reactor system asclaimed in claim 10, wherein said material capable of transferring heatinto said selected reaction volume when suitably energized is providedas concentric bands on said disc.
 12. The reactor system as claimed inclaim 11, wherein the inner and outer bands are wider than theintermediate bands.
 13. The reactor system as claimed in claim 9,wherein said material is a material capable of absorbing electromagneticradiation.
 14. The reactor system as claimed in claim 10, wherein saidchannel structure has a generally radial extension on said disc.
 15. Thereactor system as claimed in claim 10, wherein said rotatable disccomprises a base portion having a top and a bottom side, on the top sideof which said micro channel structure is provided, and on top of which acover is provided so as to seal the micro channel structure, and whereinsaid light absorbing material is provided on said bottom side or on saidtop side or on top of said cover.
 16. The reactor system as claimed inclaim 9, wherein said heating structure comprises a separate memberdisposed so as to mask electromagnetic radiation directed towards thesurface of the substrate, and having openings defining said pattern, andwherein said light absorbing material is provided over essentially theentire surface of each selected region to be heated.
 17. The reactorsystem as claimed in claim 9, wherein said material is a resistivematerial capable of generating heat when energized with electricity. 18.A heating structure for enabling the generation of a uniform temperatureprofile across a reaction volume that is part of a microchannelstructure and is present in a selected area of a substrate whichcontains said microchannel structure, said heating structure comprisinga plurality of regions of a material forming heating elements capable oftransmitting heat into said selected area when suitably energized, saidregions being provided over said selected area as a heating elementstructure defining a) the selected area to be heated, and b) the uniformtemperature profile across the reaction volume; and wherein theplurality of regions of a material is laid out in a pattern that causesheating and cooling to balance each other, so as to create said uniformtemperature profile.
 19. The heating structure as claimed in claim 18,wherein said heating elements are areas of a layer of a light absorbingmaterial.
 20. The heating structure as claimed in claim 18, wherein saidheating elements are areas of a resistive material that generates heatwhen a voltage is applied or a current is driven therethrough.
 21. Theheating structure as claimed in claim 18, wherein said heating elementsare provided as concentric bands of said light absorbing material or ofsaid resistive material, said concentric bands covering the selectedarea to be heated.
 22. The heating structure as claimed in claim 19,wherein said light absorbing material is provided on said substrate invarying thickness over said selected area, the thickness variationdefining said uniform temperature profile.
 23. The heating structure asclaimed in claim 19, wherein said light absorbing material is providedon said substrate as dots in a pattern of varying dot density, saiddensity variation defining said uniform temperature profile.
 24. Arotatable disc comprising a micro channel reactor system as claimed inclaim
 9. 25. The disc as claimed in claim 24, further comprisingrecessed portions in the substrate such that the material thickness atthe periphery of the selected regions is smaller than the nominalthickness of the substrate.
 26. The disc as claimed in claim 24, furthercomprising holes in the substrate at the periphery of the selectedregions.
 27. The method as claimed in claim 2, wherein the substrate isa rotatable disc, said pattern of said heating structure comprises amaterial capable of absorbing electromagnetic energy, and wherein heatenergy is supplied by irradiation of the disc using a source ofelectromagnetic radiation, wherein the source is a light source.
 28. Thereactor system as claimed in claim 9, wherein said substrate is arotatable disc.
 29. The method of claim 1, wherein said pattern providesthat said reaction volume is covered by one or more areas that areheated and one or more areas that are not heated.
 30. The microchannelreactor system of claim 9, wherein said pattern provides that saidreaction volume is covered by one or more areas that are heated and oneor more areas that are not heated.
 31. The heating structure of claim18, wherein said pattern provides that said reaction volume is coveredby one or more areas that are heated and one or more areas that are notheated.